Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 510838
Title Optimization of canine interleukin-12 production using a baculovirus insect cell expression system Biotechnology
Author(s) Pinheiro, Cristiane Garboggini Melo De; Pedrosa, Mayara De Oliveira; Teixeira, Naiara Carvalho; Ano Bom, Ana Paula Dinis; Oers, Monique M. Van; Sá Oliveira, Geraldo Gileno de
Source BMC Research Notes 9 (2016). - ISSN 1756-0500
DOI http://dx.doi.org/10.1186/s13104-016-1843-7
Department(s) Laboratory of Virology
PE&RC
EPS
Publication type Refereed Article in a scientific journal
Publication year 2016
Keyword(s) Baculovirus - Dog - Interleukin-12 - Protein expression optimization
Abstract

Background: Interleukin-12 is an important cytokine in mediating cellular immune responses. Results: Recombinant single-chain canine IL-12 was produced in a baculovirus-insect cell system with the aim of conducting further studies on modulation of immune responses in dogs. To optimize the production of recombinant canine IL-12, a classical baculovirus and a modified vector (chitinase A and v-cathepsin knockout) were used containing a native or an optimized insert of canine IL-12. The optimized IL-12 construct contained the GP64 signal peptide and was synthesized with optimized codons for expression in Trichoplusia ni cells. Dot-blot and Western blot analysis showed the highest production levels of recombinant IL-12 protein by the use of the modified baculovirus vector containing the optimized insert, at a multiplicity of infection of five and at 48 h after infection. The recombinant cytokine was successfully purified and showed a good degree of purity, integrity, folding, and yield, with very little endotoxin contamination. Recombinant canine IL-12 induced IFN-γ in canine lymphocytes, indicating that it was biologically active. Conclusion: Therefore, this study describes an efficient method to produce adequate amounts of biologically active canine IL-12, useful for immunomodulation studies in dogs.

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